eThe anaerobic oxidation of ammonium (anammox) process has been observed in diverse terrestrial ecosystems, while the contribution of anammox to N 2 production in paddy soils is not well documented. In this study, the anammox activity and the abundance and diversity of anammox bacteria were investigated to assess the anammox potential of 12 typical paddy soils collected in southern China. Anammox bacteria related to "Candidatus Brocadia" and "Candidatus Kuenenia" and two novel unidentified clusters were detected, with "Candidatus Brocadia" comprising 50% of the anammox population. The prevalence of the anammox was confirmed by the quantitative PCR results based on hydrazine synthase (hzsB) genes, which showed that the abundance ranged from 1.16 ؋ 10 4 to 9.65 ؋ 10 4 copies per gram of dry weight. The anammox rates measured by the isotope-pairing technique ranged from 0.27 to 5.25 nmol N per gram of soil per hour in these paddy soils, which contributed 0.6 to 15% to soil N 2 production. It is estimated that a total loss of 2.50 ؋ 10 6 Mg N per year is linked to anammox in the paddy fields in southern China, which implied that ca. 10% of the applied ammonia fertilizers is lost via the anammox process. Anammox activity was significantly correlated with the abundance of hzsB genes, soil nitrate concentration, and C/N ratio. Additionally, ammonia concentration and pH were found to be significantly correlated with the anammox bacterial structure.

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Anaerobic oxidation of ammonium (anammox) is recognized as an important process for nitrogen (N) cycling, yet its role in agricultural ecosystems, which are intensively fertilized, remains unclear. In this study, we investigated the presence, activity, functional gene abundance and role of anammox bacteria in rhizosphere and non-rhizosphere paddy soils using catalyzed reporter deposition–fluorescence in situ hybridization, isotope-tracing technique, quantitative PCR assay and 16S rRNA gene clone libraries. Results showed that rhizosphere anammox contributed to 31–41% N2 production with activities of 0.33–0.64 nmol N2 g−1 soil h−1, whereas the non-rhizosphere anammox bacteria contributed to only 2–3% N2 production with lower activities of 0.08–0.26 nmol N2 g−1 soil h−1. Higher anammox bacterial cells were observed (0.75–1.4 × 107 copies g−1 soil) in the rhizosphere, which were twofold higher compared with the non-rhizosphere soil (3.7–5.9 × 106 copies g−1 soil). Phylogenetic analysis of the anammox bacterial 16S rRNA genes indicated that two genera of ‘Candidatus Kuenenia' and ‘Candidatus Brocadia' and the family of Planctomycetaceae were identified. We suggest the rhizosphere provides a favorable niche for anammox bacteria, which are important to N cycling, but were previously largely overlooked.

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Ammonia-oxidizing archaea (AOA) and bacteria (AOB) play important roles in nitrogen cycling. However, the effects of environmental factors on the activity, abundance, and diversity of AOA and AOB and the relative contributions of these two groups to nitrification in paddy soils are not well explained. In this study, potential nitrification activity (PNA), abundance, and diversity of amoA genes from 12 paddy soils in Southern China were determined by potential nitrification assay, quantitative PCR, and cloning. The results showed that PNA was highly variable between paddy soils, ranging from 4.05 ± 0.21 to 9.81 ± 1.09 mg NOx-N kg(-1) dry soil day(-1), and no significant correlation with soil parameters was found. The abundance of AOA was predominant over AOB, indicating that AOA may be the major members in aerobic ammonia oxidation in these paddy soils. Community compositions of AOA and AOB were highly variable among samples, but the variations were best explained by pH. AOA sequences were affiliated to the Nitrosopumilus cluster and Nitrososphaera cluster, and AOB were classified into the lineages of Nitrosospira and Nitrosomonas, with Nitrosospira being predominant over Nitrosomonas, accounting for 83.6 % of the AOB community. Moreover, the majority of Nitrosomonas was determined in neutral soils. Canonical correspondence analysis (CCA) analysis further demonstrated that AOA and AOB community structures were significantly affected by pH, soil total organic carbon, total nitrogen, and C/N ratio, suggesting that these factors exert strong effects on the distribution of AOB and AOA in paddy soils in Southern China. In conclusion, our results imply that soil pH was a key explanatory variable for both AOA and AOB community structure and nitrification activity.

a b s t r a c tAnaerobic oxidation of ammonium (anammox) plays an important role in nitrogen (N) loss from agricultural systems. Recently, the rice rhizosphere was demonstrated to be a hotspot for anammox, yet the dynamics of anammox activity and the distribution of anammox bacteria in rhizosphere soil at different phenological stages of rice growth are still unknown. In this study, the activity, diversity and abundance of anammox bacteria in both rhizosphere and bulk soils were investigated over the entire rice growth season. From tillering to ripening stage, significantly higher anammox bacterial abundance was detected in rhizosphere soils compared to bulk soils. The rhizosphere soils also had significantly higher anammox rates at tillering and booting stages (0.71 and 0.32 nmol N g À1 dry soil h À1 , respectively) compared to bulk soils. The anammox rate in rhizosphere soil was positively correlated to the concentrations of NO x À (total of nitrate and nitrite) and acetate. The abundance of anammox bacteria was significantly correlated with the concentration of succinate in rhizosphere soils. A total of five anammox genera of Brocadia, Kuenenia, Anammoxoglobus, Jettenia and Scalindua were detected, with Brocadia predominating in all examined samples. The distribution of anammox bacteria in rhizosphere and bulk soils varied with phenological stages. Statistical analysis indicated that C/N ratio, formate, citrate and ammonium were key factors influencing the composition of anammox bacteria. Variations in activity, abundance and distribution of anammox bacteria in rhizosphere were observed over the phenological progression, demonstrating that the root exudates might be influential for the anammox process. This study implies that future efforts in estimating the rate of anammox should consider the temporal variation during plant life cycles.

The aim of this study was to evaluate short-term concentration and time effects of cadmium on Kandelia obovata (S., L.) Yong root exudation, thereby evaluating and predicting the ecophysiological effects of mangrove to heavy metals at the root level. Mature K. obovata propagules were cultivated in a sandy medium for 3 months, and then six concentrations of Cd (0, 2.5, 5, 10, 20, and 40 mg L(-1)) were applied. After exposure time of 24 h and 7 days, respectively, the root exudates of K. obovata were collected and low molecular weight organic acids (LMWOAs) and amino acids of which were analyzed. In addition, we measured glutathione, soluble protein content, and Cd concentration in the plant. We found 10 and 15 types of LMWOAs and amino acids in root exudates of K. obovata with total concentrations ranging from 29.54 to 43.08 mg g(-1) dry weight (DW) roots and from 737.35 to 1,452.46 ng g(-1) DW roots, respectively. Both of them varied in quality and quantity under different Cd treatment strengths and exposure times. Oxalic, acetic, L-malic, tartaric acid, tyrosine, methionine, cysteine, isoleucine, and arginine were dominant. Both LMWOAs and amino acids excreted from K. obovata roots play a key role in Cd toxicity resistance. The responsiveness of amino acids was less than that of LMWOAs. We suggest that the ecological effect of root-excreted free amino acids in the rhizosphere is mainly based on the role of nutrients, supplemented with detoxification to heavy metals.

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